Alisson Martins-Oliveira, Michele M. Castro, Diogo M. M

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Contrasting effects of aliskiren versus losartan on hypertensive vascular remodeling Alisson Martins-Oliveira, Michele M. Castro, Diogo M.M. Oliveira, Elen Rizzi, Carla S. Ceron, Danielle Guimaraes, Rosana I. Reis, Claudio M. Costa-Neto, Dulce E. Casarini, Amanda A. Ribeiro, Raquel F. Gerlach, Jose E. Tanus-Santos International Journal of Cardiology Volume 167, Issue 4, Pages 1199-1205 (August 2013) DOI: 10.1016/j.ijcard.2012.03.137 Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 Systolic blood pressure (mm Hg) measured by tail-cuff method (Panel A) and plasma renin activity (Panel B) in all experimental groups at the end of six weeks of antihypertensive treatment. Data are shown as mean±S.E.M (n=11–15 per group). *P<0.05 for the 2K1C groups versus the Sham groups. **P<0.05 versus the Sham+vehicle or versus 2K-1C+vehicle. #P<0.05 for the 2K1C group versus the other 2K1C groups. International Journal of Cardiology 2013 167, 1199-1205DOI: (10.1016/j.ijcard.2012.03.137) Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 Effects of aliskiren and/or losartan on the structural modifications induced by 2K1C hypertension in the aortas from rats. Panel A shows representative photomicrographs of aortic samples (400×) stained by hematoxylin and eosin (H&E). Panel B shows the values for vascular smooth muscle cell number per length of aorta. Panel C shows the values for thoracic aorta medial cross-sectional area (CSA). Panel D shows the values for media to lumen ratio (M/L). Data are shown as mean±S.E.M. (n=5–6 per group). *P<0.05 versus the Sham groups and versus the 2K1C+LOS and the 2K1C+ALK/LOS groups. International Journal of Cardiology 2013 167, 1199-1205DOI: (10.1016/j.ijcard.2012.03.137) Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 Effects of aliskiren and/or losartan on collagen and elastin content in the aortic media. Panels A and C show representative photomicrographs of aortic samples (400×) stained by Sirus red and Orceine, respectively. Panel B and Panel D show the values for surface area of collagen and elastin in the aortic media. Data are shown as mean±S.E.M. (n=4–6 per group). *P<0.05 versus the Sham groups and versus the 2K1C+LOS and the 2K1C+ALK/LOS groups. International Journal of Cardiology 2013 167, 1199-1205DOI: (10.1016/j.ijcard.2012.03.137) Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions

Fig. 4 A representative gelatin zymogram of aortic samples (Panel A). MMP-2 bands were identified at 75kDa, 72kDa, and 64kDa after electrophoresis on 12% SDS–PAGE. Densitometric analyses for these aortic MMP-2 forms are shown in panels B–D. Data are shown as mean±S.E.M. (n=9–11 per group). *P<0.05 versus the Sham groups and versus the other 2K1C groups. **P<0.05 versus the Sham groups only. #P<0.05 versus the Sham groups and versus the 2K1C+LOS and the 2K1C+ALK/LOS groups. International Journal of Cardiology 2013 167, 1199-1205DOI: (10.1016/j.ijcard.2012.03.137) Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions

Fig. 5 Effects of aliskiren, losartan, or aliskiren+losartan on aortic in situ gelatinolytic activity and MMP-2 levels in the aortas from Sham and 2K1C rats. Panel A shows representative photomicrographs of in situ gelatinolytic activity (×400), MMP-2 detected by immunofluorescence (M = media; L = lumen), and their co-localization (merge) in vessel surface. Panel B shows the quantification of percentage of vessel surface area covered by bright green fluorescence, which reflects gelatinolytic activity. Panel C shows the quantification of percentage of vessel surface area covered by bright red fluorescence, which reflects MMP-2 levels. Data are shown as mean±S.E.M. (n=5–6 per group). *P<0.05 versus the Sham groups and versus the 2K1C+LOS and the 2K1C+ALK/LOS groups. International Journal of Cardiology 2013 167, 1199-1205DOI: (10.1016/j.ijcard.2012.03.137) Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions

Fig. 6 Expression of MMP-2 (panel A), PRR (panel B), p-ERK-1/2 (panel C), and TGF-β1 (panel D) in the thoracic aorta from Sham and 2K1C rats after treatment with vehicle, aliskiren (ALK), losartan (LOS) or aliskiren+losartan (ALK/LOS). Densitometric intensity corresponding to each band was normalized using β-actin expression. Data are expressed as a ratio between the expression interest protein and β-actin expression. Data are shown as mean±S.E.M. (n=3–5 per group). * P<0.05 versus the Sham groups and versus the 2K1C+LOS and the 2K1C+ALK/LOS groups. International Journal of Cardiology 2013 167, 1199-1205DOI: (10.1016/j.ijcard.2012.03.137) Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions